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M-T7 modulates the immune response in the healing wound. (A) ELISA quantification of CCL2 in wounds treated with saline or M-T7 at days 1, 4 and 7 post-wounding, normalized to total protein. (B) Quantification of Arginase-1+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding. (C) Representative Arginase-1 IHC fields at day 7. (D) Quantification of TGF-beta+ cells per 20× field on days 2, 4 and 7 post-wounding. (E,F) Quantification of CD3+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding, specifically in the (E) wound bed or (F) epithelial tongue. (G) Quantification of CD4+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding normalized to the numbers on day 2. (H) Representative CD4 IHC fields in the epithelial tongue at day 7. Full 20× field is given in Figure S2. All bars are mean and standard error. Statistics are calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. N = 3-4 in each group and time point.

M-T7 modulates the immune response in the healing wound. (A) ELISA quantification of CCL2 in wounds treated with saline or M-T7 at days 1, 4 and 7 post-wounding, normalized to total protein. (B) Quantification of Arginase-1+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding. (C) Representative Arginase-1 IHC fields at day 7. (D) Quantification of TGF-beta+ cells per 20× field on days 2, 4 and 7 post-wounding. (E,F) Quantification of CD3+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding, specifically in the (E) wound bed or (F) epithelial tongue. (G) Quantification of CD4+ cells per 20× field of wounds treated with saline or M-T7 at days 2, 4 and 7 post-wounding normalized to the numbers on day 2. (H) Representative CD4 IHC fields in the epithelial tongue at day 7. Full 20× field is given in Figure S2. All bars are mean and standard error. Statistics are calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. N = 3-4 in each group and time point.

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Complex dermal wounds represent major medical and financial burdens, especially in the context of comorbidities such as diabetes, infection and advanced age. New approaches to accelerate and improve, or "fine tune" the healing process, so as to improve the quality of cutaneous wound healing and management, are the focus of intense investigation. He...

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Context 1
... also called monocyte chemoattractant protein-1 (MCP-1), is a CC-class chemokine previously shown to have a critical role in the regulation of wound healing [48]. We performed the ELISA analysis of CCL2 on wound tissues treated with saline or M-T7, collected on days 1, 4 and 7 post-wounding ( Figure 4A). Wounds treated with M-T7 had an elevated level of CCL2 on day 4 post-wounding, which approached significance (p = 0.0763), while levels of CCL2 were not different between saline and M-T7 treatment on days 1 and 7 post-wounding. ...
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... performed immunohistochemistry of wounds treated with saline or M-T7 on days 2, 4 and 7 post-wounding, staining for Arginase-1, a canonical marker of M2 macrophage polarization. The quantification of Arginase-1+ cells revealed a trend towards elevated M2 macrophages on days 2 and 4 post-wounding, which achieved significance (p < 0.05) by day 7 post-wounding ( Figure 4B,C). Accordingly, the number of TGF-beta+ cells per field trended towards significance on day 4 (p = 0.0836) and reached significance by day 7 post-wounding (p < 0.05) ( Figure 4D). ...
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... quantification of Arginase-1+ cells revealed a trend towards elevated M2 macrophages on days 2 and 4 post-wounding, which achieved significance (p < 0.05) by day 7 post-wounding ( Figure 4B,C). Accordingly, the number of TGF-beta+ cells per field trended towards significance on day 4 (p = 0.0836) and reached significance by day 7 post-wounding (p < 0.05) ( Figure 4D). We further investigated the effects of M-T7 treatment on T cell infiltration in the healing wound. ...
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... further investigated the effects of M-T7 treatment on T cell infiltration in the healing wound. We found that M-T7 treatment significantly inhibited the infiltration of CD3+ T cells, a general T cell marker, into the bed of the healing wound on days 4 and 7 post-wounding ( Figure 4E), without inhibiting the accumulation of CD3+ cells in the epithelial tongue of the wounds ( Figure 4F). Regulatory T cells, a CD4 T cell subtype, are crucial for the normal and accelerated healing of cutaneous wounds [50]. ...
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... further investigated the effects of M-T7 treatment on T cell infiltration in the healing wound. We found that M-T7 treatment significantly inhibited the infiltration of CD3+ T cells, a general T cell marker, into the bed of the healing wound on days 4 and 7 post-wounding ( Figure 4E), without inhibiting the accumulation of CD3+ cells in the epithelial tongue of the wounds ( Figure 4F). Regulatory T cells, a CD4 T cell subtype, are crucial for the normal and accelerated healing of cutaneous wounds [50]. ...
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... the dimerization of CCL2 requires glycosaminoglycan interactions [71]. We found increased levels of CCL2 in healing wounds when treated with recombinant M-T7 ( Figure 4A), consistent with the role of CCL2 in improved healing. We hypothesize that M-T7 treatment inhibited the oligomerization of CCL2, slowing receptor engagement and delaying its subsequent degradation. ...
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... further, we found M-T7-dependent effects in two cell populations known to be affected by CCL2 and other chemokine signaling: macrophages and T cells. Specifically, we found an increase in M2-polarized, proresolution macrophages ( Figure 4B). This finding agrees with prior work showing that CCL2 signaling results in M2 polarization [49]. ...
Context 8
... finding agrees with prior work showing that CCL2 signaling results in M2 polarization [49]. We also found increased CD4+ T cells in the epithelial tongues of healing wounds treated with M-T7 ( Figure 4F,G), in agreement with the ability for CCL2 to promote CD4 recruitment, and in the CD4-lineage cells driving accelerated wound healing [50,72]. CCL2 acts directly on T cells via the action of CCR2 and CCR4 [73], but can also induce the recruitment of CD4 cells into tissues in a promiscuous manner, using other receptors [72]. ...

Citations

... The splinted full thickness model was used study the effects of recombinant myxoma virus-derived immune modulator M-T7 [49], of Babassu oil [50], of three-dimensional printed scaffolds and electrospun mats [51], of adult gingival multipotent mensenchymal stem cells [52], of induced pluripotent stem cells clay [53], of poly(lactic-co-glycolic acid) and vascular endothelial growth factor [54], and of renal dysfunction [44] on cutaneous wound healing. ...
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A large number of models are now available for the investigation of skin wound healing. These can be used to study the processes that take place in a phase-specific manner under both physiological and pathological conditions. Most models focus on wound closure, which is a crucial parameter for wound healing. However, vascular supply plays an equally important role and corresponding models for selective or parallel investigation of microcirculation regeneration and angiogenesis are also described. In this review article, we therefore focus on the different levels of investigation of skin wound healing (in vivo to in virtuo) and the investigation of angiogenesis and its parameters.
... This finding is entirely consistent with prior research both in pre-clinical or clinical studies wherein Serp-1 demonstrated no significant toxicity and no neutralizing antibodies were detected in the Phase Further studies should also aim to investigate the molecular mechanism of action of Serp-1 in its anti-inflammatory role. Finally, a recent publication from our group demonstrated that administration of recombinant M-T7 protein (another Myxoma virus protein) could also accelerate dermal wound healing (24). This provides an additional example of a Myxoma virus derived protein in facilitating tissue repair. ...
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